Heat treatment to produce glycosides

Patent metadata

Field	Value
Publication number	US-11028423-B2
Application number	US-202016796630-A
Country	US
Kind code	B2
Filing date	Feb 20, 2020
Priority date	May 29, 2015
Publication date	Jun 8, 2021
Grant date	Jun 8, 2021

Abstract

Disclosed arc methods of releasing or enriching for steviol glycosides produced by yeast. The disclosed methods enhance or improve the release or enrichment of the steviol glycosides.

First claim

The invention claimed is:

1. A method of releasing steviol glycosides from a yeast host cell, comprising the steps of:
(a) providing a composition, comprising the host cell producing one or more steviol glycosides; and
(b) releasing the one or more steviol glycosides from the host cell by heating the composition to a temperature in a range from 50° C. to 95° C. for 5 minutes to 48 hours, wherein the one or more steviol glycosides are released from an intracellular portion of the host cell into an extracellular portion of the composition.

2. The method of claim 1 , wherein the composition is a liquid composition comprising the host cell and a liquid media.

3. The method of claim 2 , wherein the composition is obtained by separating the host cell from a fermentation medium comprising the host cell, and transferring the separated host cell into the suitable liquid media.

4. The method of claim 1 , wherein the composition is a fermentation medium.

5. The method of claim 1 , wherein the host cell is concentrated before the heat treatment.

6. The method of claim 1 , wherein the composition is heated to the temperature in the range from 70° C. to 95° C. for 5 minutes to 2 hours.

7. The method of claim 1 , wherein the composition is heated to about 50° C. for 24 hours.

8. The method of claim 1 , wherein the composition is heated to about 70° C. for 5 minutes to 2 hours.

9. The method of claim 1 , wherein the composition is heated to about 75° C. for 5 minutes to 2 hours.

10. The method of claim 1 , wherein the composition is heated to about 95° C. for 5 minutes to 1 hour.

11. The method of claim 1 , wherein an amount of the steviol glycosides released extracellularly is in a range of about 80% to about 100% of a total intracellular and extracellular amount of the steviol glycosides produced by the host cell.

12. The method of claim 1 , wherein the one or more steviol glycosides is rebaudioside M (Reb M), rebaudioside D (Reb D), rebaudioside A (Reb A), rebaudioside B (Reb B), or combinations thereof.

13. The method of claim 12 , wherein an amount of Reb A released extracellularly is in a range of about 30% to about 100% of a total intracellular and extracellular amount of Reb A produced by the host cell.

14. The method of claim 12 , wherein an amount of Reb B released extracellularly is in a range of about 25% to about 100% of a total intracellular and extracellular amount of Reb B produced by the host cell.

15. The method of claim 12 , wherein an amount of Reb D and/or rebaudioside M released extracellularly is in a range of about 30% to about 100% of a total intracellular and extracellular amount of Reb D and/or Reb M produced by the host cell.

16. The method of claim 1 , wherein an extracellular nitrogen content in the composition after the heat treatment is not increased by more than 50% of a total extracellular and intracellular nitrogen content in the composition.

17. The method of claim 1 , wherein an extracellular phosphorous content in the composition after the heat treatment is not increased by more than 80% of a total extracellular and intracellular phosphorous content in the composition.

18. The method of claim 1 , wherein the yeast is selected from a group consisting of genus, Candida, Pichia ( Hansenula ), Kloeckera ( Hanseniaspora ), Kluyveromyces, Rhodotorula, Torulopsis, Zygosaccharomyces, Saccharomycete, Yarrowia , and Saccharomyces.

19. The method of claim 1 , wherein the yeast is Saccharomyces cerevisiae.

20. The method of claim 1 , wherein the released steviol glycosides are further isolated or purified from the host cell.

21. A composition, comprising a yeast host cell producing one or more steviol glycosides and one or more steviol glycosides that are released from the host cell by heating the composition to a temperature in a range from 50° C. to 95° C. for 5 minutes to 48 hours;
wherein an amount of the steviol glycosides released extracellularly is in a range of about 80% to about 100% of a total intracellular and extracellular amount of the steviol glycosides produced by the host cell.

Assignees

• Cargill Inc

Inventors

• Anderson James C
• Brower Iii Robert J
• Carlson Ting Liu
• Flores Belit
• Gaspard Dan S
• Mortenson Kristopher T
• Nygaard Richard
• Paulson Nicole
• Rasmussen Maribeth

Classifications

• C12N15/81

  for yeasts · CPC title

• A61K31/704

  attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin {(digitoxin A61K31/7048)} · CPC title

• C12N2523/00

  Culture process characterised by temperature · CPC title

• A23L2/60

  Sweeteners · CPC title

• C12P19/56 Primary

  having an oxygen atom of the saccharide radical directly bound to a condensed ring system having three or more carbocyclic rings, e.g. daunomycin, adriamycin · CPC title